NextSeq 550 sequencing; Flexible wood caused by altered lignin, following viral-induced suppression of phenylalanine ammonia lyase (PAL) activity. sRNA-seq
Flexible wood caused by altered lignin, following viral-induced suppression of phenylalanine ammonia lyase (PAL) activity. sRNA-seq
RNA was extracted from 10 mg of stem material. 6 uninfected and 6 infected graft segments were used. After removing the bark, whole xylem segments were flash frozen in liquid nitrogen. Newly differentiating xylem was scraped from frozen xylem segments and then ground into a fine powder using a mortar and pestle with liquid nitrogen. Apple stem material was propagated from 2 uninfected and 2 ARW-infected Lord Lambourne apple trees, which was provided by the East Malling Research Station, Kent, UK in 1996. Budwood was grafted onto M27 rootstocks using the 'whip and tongue' grafting method (Bradley & Garner, 2013) in April 2017. Grafts were kept in a glass house at 22°C and one-year old stem material was harvested the following year for analysis. Qiagen RNAeasy Plant Power kit to extract RNA from ground xylem samples. DNA was removed with the Qiagen DNAse Max kit. NEBNext Small RNA Library Prep